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Characterisation of the Brochothrix thermosphacta sortase A enzyme
Stanborough, Tamsyn and Suryadinata, R and Fegan, N and Powell, SM and Tamplin, M and Nuttall, SD and Chandry, PS, Characterisation of the Brochothrix thermosphacta sortase A enzyme, FEMS Microbiology Letters, 365, (17) pp. 1-9. ISSN 1574-6968 (2018) [Refereed Article]
© FEMS 2018.
Official URL: https://academic.oup.com/femsle/article/365/17/fny...
Gram-positive bacteria utilise class A sortases to coat the surface of their cells with a diversity of proteins that facilitate interactions with their environment and play fundamental roles in cell physiology and virulence. A putative sortase A gene was identified in the genome of the poorly studied meat spoilage bacterium Brochothrix thermosphacta. To understand how this bacterium mediates interactions with its environment, an N-terminal truncated, His-tagged variant of this protein (His6-BtSrtA) was expressed and purified. Catalytic activity of recombinant His6-BtSrtA was investigated, including sorting motif recognition of target proteins and bioconjugation activity. Further, the B. thermosphacta genome was examined for the presence of sortase A (SrtA) protein substrates. His6-BtSrtA readily formed intermediate complexes with LPXTG-tagged proteins. Although the reaction was inefficient, nucleophilic attack of the resultant thioacyl intermediates by tri-glycine was observed. Genome examination identified 11 potential SrtA substrates, two of which contained protein domains associated with adherence of pathogens to host extracellular matrix proteins and cells, suggesting the B. thermosphacta SrtA may be indirectly involved in its attachment to meat surfaces. Thus, further work in this area could provide crucial insight into molecular mechanisms involved in the colonisation of meat by B. thermosphacta.
|Item Type:||Refereed Article|
|Research Division:||Biological Sciences|
|Objective Group:||Public health (excl. specific population health)|
|Objective Field:||Food safety|
|UTAS Author:||Stanborough, Tamsyn (Ms Tamsyn Stanborough)|
|UTAS Author:||Fegan, N (Ms Narelle Fegan)|
|UTAS Author:||Powell, SM (Dr Shane Powell)|
|UTAS Author:||Tamplin, M (Professor Mark Tamplin)|
|UTAS Author:||Chandry, PS (Dr Peter Chandry)|
|Web of Science® Times Cited:||2|
|Deposited By:||Agriculture and Food Systems|
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